MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT CGRP RECEPTOR
1 vial (2 x 106) frozen cells
Cell Banker 2 (Amsbio 11891)
Full-length Human CALCRL cDNA (GenBank Accession Number ( NM_005795) with FLAG-tag sequence at the N-terminus and Full- length Human receptor activity modifying protein 1 (RAMP1) cDNA (GenBank Accession Number BC000548) with myc-tag at the C- terminus
Liquid nitrogen upon receiving
Propagation Medium: DME/F12, 10% FBS, 10 µg/mL puromycin, 250µg/mL hygromycin
Stable in culture for minimum of two months
Background: CALCRL (calcitonin receptor-like receptor) is a calcitonin family receptor. The function and pharmacology of this receptor is modified in the presence of receptor activity-modifying proteins (RAMPs), which are single Transmembrane domain proteins of about 160 amino acids, having three isoforms: RAMP1, RAMP2 and RAMP3. CALCR can function either as a calcitonin gene-related peptide (CGRP) receptor or an adrenomedullin (AM) receptor, depending on which members of RAMPs, are co-expressed. The CALCRL/RAMP1 complex has been known as the CGRP receptor and has a very high affinity for CGRP.
Application: Functional assays
Figure 1. Dose-dependent increase of intracellular cAMP level upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 2. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Morfis et al. (2008) Receptor Activity-Modifying Proteins Differentially Modulate the G Protein-coupling Efficiency of Amylin Receptors. Endocrinology: 149(11):5423–5431.
Hay et al. (2005) Pharmacological Discrimination of Calcitonin Receptor: Receptor Activity-Modifying Protein Complexes. Mol Pharmacol 67:1655–1665.
Gorn et al. (1992) Cloning, characterization, and expression of a human calcitonin receptor from an ovarian carcinoma cell line. J Clin Invest 90:1726-1735.
RJ Bailey et al.(2011). Pharmacological characterization of rat amylin receptors: implications for the identification of amylin receptor subtypes. BJP 166:151–167